Background Pre-hospital emergency care is a medical care given to patients before arrival in the hospital after activation of the emergency team. Poor knowledge and practice about pre-hospital emergency care hurt the health outcomes of the patients. Objective This study aimed to assess knowledge and practice nurses at the University of Gondar Compressive Specialized Hospital, Northwest Ethiopia. Methods An institutional-based cross-sectional study was conducted from March 20 to April 10, 2020. A stratified sampling technique was used to select the study participants. Data were collected using a pretested structured self-administered questionnaire. Data were analyzed using SPSS version 20. To explain study variables, frequency tables and percentages were used. Logistic regression analysis was used to see the association between independent and dependent variables. Results Out of the total 378 respondents, less than half (42.9%) had good knowledge; similarly, 49.5% of them had good practice about pre-hospital emergency care. Male sex and attend formal training were significant associations with both knowledge and practice of pre-hospital emergency nursing care. Male participants (adjusted odds ratio (AOR) = 6.57, 95% confidence interval (CI) (3.79–11.36)) and having training (AOR=1.74, 95% CI (1.83–3.66)) were significantly associated with knowledge of pre-hospital emergency care, whereas male sex (AOR=1.73, 95% CI (1.09–2.73)) and having training (AOR=6.16, 95% CI (2.69–14.10)) were significantly associated with the practice of pre-hospital emergency care. Conclusion Knowledge and practice of nurses regarding pre-hospital emergency care was found to be inadequate as compared to previous studies. Male sex and attend formal training showed a positive and significant association with both knowledge and practice of pre-hospital emergency nursing care. The responsible body ought to allow professional development and attending formal training for nurses.